US2024335539A1PendingUtilityA1
Modulation of bcl-2 to enhance chimeric antigen receptor cancer immunotherapy efficacy

Assignee: UNIV PENNSYLVANIAPriority: Aug 11, 2021Filed: Aug 10, 2022Published: Oct 10, 2024
Est. expiryAug 11, 2041(~15 yrs left)· nominal 20-yr term from priority
A61K 40/4211A61K 40/31A61K 40/11A61K 40/421A61K 40/30A61K 2239/38A61K 2239/48C12N 2740/15043C12N 15/86C07K 2319/30C07K 2319/02C07K 2317/622C07K 16/2803C07K 14/7051C07K 14/4747A61P 35/00C07K 2319/92C07K 2319/03C12N 2510/00C12N 5/0636C07K 14/82C12N 15/62C07K 2319/50A61K 39/4637A61K 39/4631A61K 39/464412
PatentIndex Score
Cited by
References
Claims
Abstract

The present disclosure provides modified cell(s), i.e., immune cell(s) or precursor cell(s) thereof, wherein the cell(s) are engineered to express a) a chimeric antigen receptor (CAR), and b) a variant of a B-cell lymphoma 2 (Bcl-2) family protein, wherein the variant confers resistance to a cytotoxic inhibitor of the Bcl-2 family protein. Also provided are methods and uses of the modified cells, e.g., for treating at least one sign and/or symptom of cancer. Related nucleic acids, vectors, and pharmaceutical compositions are also provided.
Claims

exact text as granted — not AI-modified
1 . An isolated nucleic acid comprising:
 a. a nucleotide sequence encoding a chimeric antigen receptor (CAR) comprising an extracellular antigen binding domain, a transmembrane domain, and an intracellular domain, wherein the antigen binding domain binds a tumor antigen; and   b. a nucleotide sequence encoding a variant of a B-cell lymphoma 2 (Bcl-2) family protein, wherein the variant confers resistance to a cytotoxic inhibitor of the Bcl-2 family protein.   
     
     
         2 . The isolated nucleic acid of  claim 1 , further comprising a nucleotide sequence encoding a 2A self-cleaving peptide between the nucleotide sequence encoding a CAR and the nucleotide sequence encoding a variant of a Bcl-2 family protein. 
     
     
         3 . The isolated nucleic acid of  claim 1 , wherein the cytotoxic inhibitor is a pro-apoptotic drug. 
     
     
         4 . The isolated nucleic acid of  claim 1 , wherein the Bcl-2 family protein is selected from Bcl-2, BCL-XL, BCL-W, MCL1, BFL1, BIM, BAD, BAK, and BAX. 
     
     
         5 . The isolated nucleic acid of  claim 1 , wherein the Bcl-2 family protein is human Bcl-2 or human BAX. 
     
     
         6 . The isolated nucleic acid of  claim 1 , wherein the cytotoxic inhibitor is selected from the group consisting of a small molecule, an antibody, and an inhibitory nucleic acid. 
     
     
         7 . The isolated nucleic acid of  claim 1 , wherein the cytotoxic inhibitor is a small molecule. 
     
     
         8 . The isolated nucleic acid of  claim 1 , wherein the cytotoxic inhibitor is selected from the group consisting of venetoclax (ABT-199), navitoclax (ABT-263), ABT-737, sabutoclax (BI-97C1), obatoclax (GX15-070), TW-37, AT-101, HA14-1, RU486, BAM7, A-1331852, A-1155463, BDA-366, UMI-77, BH3I-1, and any combination thereof. 
     
     
         9 . The isolated nucleic acid of  claim 1 , wherein the cytotoxic inhibitor is venetoclax. 
     
     
         10 . The isolated nucleic acid of  claim 1 , wherein:
 a. the Bcl-2 family protein is human Bcl-2 and the variant comprises a mutation selected from the group consisting of F104L, G101V, D103E, D103Y, F101C, F101L, V92L, T187I, A131V, and any combination thereof, or   b. the Bcl-2 family protein is human BAX and the variant comprises a G179E mutation.   
     
     
         11 . The isolated nucleic acid of  claim 1 , wherein the variant comprises F104L Bcl-2. 
     
     
         12 . The isolated nucleic acid of  claim 1 , wherein the tumor antigen is selected from the group consisting of alpha feto-protein (AFP)/HLA-A2, AXL, B7-H3, BCMA, CA-1X, CD2, CD3, CD4, CD5, CD7, CD8, CD19, CD20, CD22, CD30, CD33, CD38, CD44v6, CD70, CD79a, CD79b, CD80, CD86, CD117, CD123, CD133, CD147, CD171, CD276, CEA, claudin 18.2, c-Met, DLL3, DR5, EGFR, EGFRvIII, EpCAM, EphA2, FAP, folate receptor alpha (FRa)/folate binding protein (FBP), GD-2, Glycolipid F77, glypican-3 (GPC3), HER2, HLA-A2, ICAM1, IL3Ra, IL13Ra2, LAGE-1, Lewis Y, LMP1 (EBV), MAGE-A1, MAGE-A3, MAGE-A4, Melan A, mesothelin, MG7 (glycosylated CEA), MMP, MUC1, Nectin4/FAP, NKG2D-Ligands (MIC-A, MIC-B, and the ULBPs 1 to 6), NY-ESO-1, P16, PD-L1, PSCA, PSMA, ROR1, ROR2, TIM-3, TM4SF1, VEGFR2, and any combination thereof. 
     
     
         13 . The isolated nucleic acid of  claim 1 , wherein the tumor antigen is CD19. 
     
     
         14 . The isolated nucleic acid of  claim 1 , wherein the antigen binding domain is selected from the group consisting of a full length antibody or antigen-binding fragment thereof, a monospecific antibody, a bispecific antibody, an Fab, an Fab′, an F(ab′)2, an Fv, a single-chain variable fragment (scFv), a linear antibody, a single-domain antibody (sdAb) and an antibody mimetic (such as a designed ankyrin repeat protein (DARPin), an affibody, a monobody (adnectin), an affilin, an affimer, an affitin, an alphabody, an avimer, a Kunitz domain peptide, an anticalin, and a syntherin). 
     
     
         15 . The isolated nucleic acid of  claim 1 , wherein the antigen binding domain is a single-chain variable fragment (scFv). 
     
     
         16 . The isolated nucleic acid of  claim 1 , wherein the intracellular domain comprises a costimulatory domain and an intracellular signaling domain. 
     
     
         17 . The isolated nucleic acid of  claim 1 , wherein the intracellular domain comprises a costimulatory domain of a protein selected from the group consisting of proteins in the TNFR superfamily, CD28, 4-1BB (CD137), OX40 (CD134), PD-1, CD7, LIGHT, CD83L, DAP10, DAP12, CD27, CD2, CD5, ICAM-1, LFA-1, Lck, TNFR-I, TNFR-II, Fas, CD30, CD40, ICOS, NKG2C, and B7-H3 (CD276), or a variant thereof, or an intracellular domain derived from a killer immunoglobulin-like receptor (KIR). 
     
     
         18 . The isolated nucleic acid of  claim 1 , wherein the intracellular domain comprises an intracellular signaling domain of a protein selected from the group consisting of a human CD3 zeta chain (CD3ζ), FcγRIII, FcsRI, DAP10, DAP12, a cytoplasmic tail of an Fc receptor, an immunoreceptor tyrosine-based activation motif (ITAM) bearing cytoplasmic receptor, TCR zeta, FcR gamma, CD3 gamma, CD3 delta, CD3 epsilon, CD5, CD22, CD79a, CD79b, and CD66d, or a variant thereof. 
     
     
         19 . The isolated nucleic acid of  claim 1 , wherein the intracellular signaling domain comprises an intracellular signaling domain of CD3ζ or a variant thereof. 
     
     
         20 . A vector comprising the isolated nucleic acid of  claim 1 . 
     
     
         21 . The vector of  claim 20 , wherein the vector is a lentiviral vector. 
     
     
         22 . A modified cell comprising the isolated nucleic acid of  claim 1 , wherein the cell is an immune cell or precursor cell thereof. 
     
     
         23 . The modified cell of  claim 22 , wherein the cell is a T cell, an autologous cell, a human cell, or any combination thereof. 
     
     
         24 . A modified cell, wherein the cell is an immune cell or precursor cell thereof, and wherein the cell is engineered to express:
 a. a chimeric antigen receptor (CAR) comprising an extracellular antigen binding domain, a transmembrane domain, and an intracellular domain, wherein the antigen binding domain binds a tumor antigen; and   b. a variant of a B-cell lymphoma 2 (Bcl-2) family protein, wherein the variant confers resistance to a cytotoxic inhibitor of the Bcl-2 family protein.   
     
     
         25 . The modified cell of  claim 24 , wherein the cytotoxic inhibitor is a pro-apoptotic drug. 
     
     
         26 . The modified cell of  claim 24 , wherein the Bcl-2 family protein is selected from Bcl-2, BCL-XL, BCL-W, MCL1, BFL1, BIM, BAD, BAK, and BAX. 
     
     
         27 . The modified cell of  claim 24 , wherein the Bcl-2 family protein is human Bcl-2 or human BAX. 
     
     
         28 . The modified cell of  claim 24 , wherein the cytotoxic inhibitor is selected from the group consisting of a small molecule, an antibody, and an inhibitory nucleic acid. 
     
     
         29 . The modified cell of  claim 24 , wherein the cytotoxic inhibitor is a small molecule. 
     
     
         30 . The modified cell of  claim 24 , wherein the cytotoxic inhibitor is selected from the group consisting of venetoclax (ABT-199), navitoclax (ABT-263), ABT-737, sabutoclax (BI-97C1), obatoclax (GX15-070), TW-37, AT-101, HA14-1, RU486, BAM7, A-1331852, A-1155463, BDA-366, UMI-77, BH3I-1, and any combination thereof. 
     
     
         31 . The modified cell of  claim 24 , wherein the cytotoxic inhibitor is venetoclax. 
     
     
         32 . The modified cell of  claim 24 , wherein:
 a. the Bcl-2 family protein is human Bcl-2 and the variant comprises a mutation selected from the group consisting of F104L, G101V, D103E, D103Y, F101C, F101L, V92L, T187I, A131V, and any combination thereof, or   b. the Bcl-2 family protein is human BAX and the variant comprises a G179E mutation.   
     
     
         33 . The modified cell of  claim 24 , wherein the variant comprises F104L Bcl-2. 
     
     
         34 . The modified cell of  claim 24 , wherein the tumor antigen is selected from the group consisting of alpha feto-protein (AFP)/HLA-A2, AXL, B7-H3, BCMA, CA-1X, CD2, CD3, CD4, CD5, CD7, CD8, CD19, CD20, CD22, CD30, CD33, CD38, CD44v6, CD70, CD79a, CD79b, CD80, CD86, CD 117, CD123, CD133, CD147, CD171, CD276, CEA, claudin 18.2, c-Met, DLL3, DR5, EGFR, EGFRvIII, EpCAM, EphA2, FAP, folate receptor alpha (FRa)/folate binding protein (FBP), GD-2, Glycolipid F77, glypican-3 (GPC3), HER2, HLA-A2, ICAM1, IL3Ra, IL13Ra2, LAGE-1, Lewis Y, LMP1 (EBV), MAGE-A1, MAGE-A3, MAGE-A4, Melan A, mesothelin, MG7 (glycosylated CEA), MMP, MUC1, Nectin4/FAP, NKG2D-Ligands (MIC-A, MIC-B, and the ULBPs 1 to 6), NY-ESO-1, P16, PD-L1, PSCA, PSMA, ROR1, ROR2, TIM-3, TM4SF1, VEGFR2, and any combination thereof. 
     
     
         35 . The modified cell of  claim 24 , wherein the tumor antigen is CD19. 
     
     
         36 . The modified cell of  claim 24 , wherein the antigen binding domain is selected from the group consisting of a full length antibody or antigen-binding fragment thereof, a monospecific antibody, a bispecific antibody, an Fab, an Fab′, an F(ab′) 2 , an Fv, a single-chain variable fragment (scFv), a linear antibody, a single-domain antibody (sdAb), and an antibody mimetic (such as a designed ankyrin repeat protein (DARPin), an affibody, a monobody (adnectin), an affilin, an affimer, an affitin, an alphabody, an avimer, a Kunitz domain peptide, an anticalin, and a syntherin). 
     
     
         37 . The modified cell of  claim 24 , wherein the antigen binding domain is a single-chain variable fragment (scFv). 
     
     
         38 . The modified cell of  claim 24 , wherein the intracellular domain comprises a costimulatory domain and an intracellular signaling domain. 
     
     
         39 . The modified cell of  claim 24 , wherein the intracellular domain comprises a costimulatory domain of a protein selected from the group consisting of proteins in the TNFR superfamily, CD28, 4-1BB (CD137), OX40 (CD134), PD-1, CD7, LIGHT, CD83L, DAP10, DAP12, CD27, CD2, CD5, ICAM-1, LFA-1, Lck, TNFR-I, TNFR-II, Fas, CD30, CD40, ICOS, NKG2C, and B7-H3 (CD276), or a variant thereof, or an intracellular domain derived from a killer immunoglobulin-like receptor (KIR). 
     
     
         40 . The modified cell of  claim 24 , wherein the intracellular domain comprises an intracellular signaling domain of a protein selected from the group consisting of a human CD3 zeta chain (CD3ζ), FcγRIII, FcsRI, DAP10, DAP12, a cytoplasmic tail of an Fc receptor, an immunoreceptor tyrosine-based activation motif (ITAM) bearing cytoplasmic receptor, TCR zeta, FcR gamma, CD3 gamma, CD3 delta, CD3 epsilon, CD5, CD22, CD79a, CD79b, and CD66d, or a variant thereof. 
     
     
         41 . The modified cell of  claim 24 , wherein the intracellular signaling domain comprises an intracellular signaling domain of CD3ζ or a variant thereof. 
     
     
         42 . The modified cell of  claim 24 , wherein the cell is a T cell, an autologous cell, a human cell, or any combination thereof. 
     
     
         43 . A pharmaceutical composition comprising a population of the modified cell of  claim 24  and at least one pharmaceutically acceptable carrier. 
     
     
         44 . A method of treating cancer in a subject in need thereof, the method comprising administering to the subject a population of modified cells, wherein the cells are immune cells or precursor cells thereof, and wherein the cells are engineered to express:
 a. a chimeric antigen receptor (CAR) comprising an extracellular antigen binding domain, a transmembrane domain, and an intracellular domain, wherein the antigen binding domain binds a tumor antigen expressed by the cancer; and   b. a variant of a B-cell lymphoma 2 (Bcl-2) family protein, wherein the variant confers resistance to a cytotoxic inhibitor of the Bcl-2 family protein.   
     
     
         45 . The method of  claim 44 , wherein the subject has been administered the cytotoxic inhibitor prior to the administration of the population of modified cells. 
     
     
         46 . The method of  claim 44 , further comprising administering the cytotoxic inhibitor to the subject prior to, simultaneously with, or after administering the population of modified cells. 
     
     
         47 . The method of  claim 44 , wherein the cytotoxic inhibitor is a pro-apoptotic drug. 
     
     
         48 . The method of  claim 44 , wherein the Bcl-2 family protein is selected from Bcl-2, BCL-XL, BCL-W, MCL1, BFL1, BIM, BAD, BAK, and BAX. 
     
     
         49 . The method of  claim 44 , wherein the Bcl-2 family protein is human Bcl-2 or human BAX. 
     
     
         50 . The method of  claim 44 , wherein the cytotoxic inhibitor is selected from the group consisting of a small molecule, an antibody, and an inhibitory nucleic acid. 
     
     
         51 . The method of  claim 44 , wherein the cytotoxic inhibitor is a small molecule. 
     
     
         52 . The method of  claim 44 , wherein the cytotoxic inhibitor is selected from the group consisting of venetoclax (ABT-199), navitoclax (ABT-263), ABT-737, sabutoclax (BI-97C1), obatoclax (GX15-070), TW-37, AT-101, HA14-1, RU486, BAM7, A-1331852, A-1155463, BDA-366, UMI-77, BH3I-1, and any combination thereof. 
     
     
         53 . The method of  claim 44 , wherein the cytotoxic inhibitor is venetoclax. 
     
     
         54 . The method of  claim 44 , wherein:
 a. the Bcl-2 family protein is human Bcl-2 and the variant comprises a mutation selected from the group consisting of F104L, G101V, D103E, D103Y, F101C, F101L, V92L, T187I, A131V, and any combination thereof, or   b. the Bcl-2 family protein is human BAX and the variant comprises a G179E mutation.   
     
     
         55 . The method of  claim 44 , wherein the variant comprises F104L Bcl-2. 
     
     
         56 . The method of  claim 44 , wherein the antigen binding domain is selected from the group consisting of a full length antibody or antigen-binding fragment thereof, a monospecific antibody, a bispecific antibody, an Fab, an Fab′, an F(ab′) 2 , an Fv, a single-chain variable fragment (scFv), a linear antibody, a single-domain antibody (sdAb), and an antibody mimetic (such as a designed ankyrin repeat protein (DARPin), an affibody, a monobody (adnectin), an affilin, an affimer, an affitin, an alphabody, an avimer, a Kunitz domain peptide, an anticalin, and a syntherin. 
     
     
         57 . The method of  claim 44 , wherein the antigen binding domain is a single-chain variable fragment (scFv). 
     
     
         58 . The method of  claim 44 , wherein the tumor antigen is selected from the group consisting of alpha feto-protein (AFP)/HLA-A2, AXL, B7-H3, BCMA, CA-1X, CD2, CD3, CD4, CD5, CD7, CD8, CD19, CD20, CD22, CD30, CD33, CD38, CD44v6, CD70, CD79a, CD79b, CD80, CD86, CD 117, CD123, CD133, CD147, CD171, CD276, CEA, claudin 18.2, c-Met, DLL3, DR5, EGFR, EGFRvIII, EpCAM, EphA2, FAP, folate receptor alpha (FRa)/folate binding protein (FBP), GD-2, Glycolipid F77, glypican-3 (GPC3), HER2, HLA-A2, ICAM1, IL3Ra, IL13Ra2, LAGE-1, Lewis Y, LMP1 (EBV), MAGE-A1, MAGE-A3, MAGE-A4, Melan A, mesothelin, MG7 (glycosylated CEA), MMP, MUC1, Nectin4/FAP, NKG2D-Ligands (MIC-A, MIC-B, and the ULBPs 1 to 6), NY-ESO-1, P16, PD-L1, PSCA, PSMA, ROR1, ROR2, TIM-3, TM4SF1, VEGFR2, and any combination thereof. 
     
     
         59 . The method of  claim 44 , wherein the tumor antigen is CD19. 
     
     
         60 . The method of  claim 44 , wherein the intracellular domain comprises a costimulatory domain and an intracellular signaling domain. 
     
     
         61 . The method of  claim 44 , wherein the intracellular domain comprises a costimulatory domain of a protein selected from the group consisting of proteins in the TNFR superfamily, CD28, 4-1BB (CD137), OX40 (CD134), PD-1, CD7, LIGHT, CD83L, DAP10, DAP12, CD27, CD2, CD5, ICAM-1, LFA-1, Lck, TNFR-I, TNFR-II, Fas, CD30, CD40, ICOS, NKG2C, and B7-H3 (CD276), or a variant thereof, or an intracellular domain derived from a killer immunoglobulin-like receptor (KIR). 
     
     
         62 . The method of  claim 44 , wherein the intracellular domain comprises an intracellular signaling domain of a protein selected from the group consisting of a human CD3 zeta chain (CD3ζ), FcγRIII, FcsRI, DAP10, DAP12, a cytoplasmic tail of an Fc receptor, an immunoreceptor tyrosine-based activation motif (ITAM) bearing cytoplasmic receptor, TCR zeta, FcR gamma, CD3 gamma, CD3 delta, CD3 epsilon, CD5, CD22, CD79a, CD79b, and CD66d, or a variant thereof. 
     
     
         63 . The method of  claim 44 , wherein the intracellular signaling domain comprises an intracellular signaling domain of CD3ζ or a variant thereof. 
     
     
         64 . The method of  claim 44 , wherein the population of cells comprises T cells, autologous cells, human cells, or any combination thereof. 
     
     
         65 . The method of  claim 44 , wherein the subject is human. 
     
     
         66 . The method of  claim 44 , wherein the cancer is B-cell lymphoma or leukemia.
Join the waitlist — get patent alerts

Track US2024335539A1 — get alerts on status changes and closely related new filings.
We store only your email — no account needed. See our privacy policy.